Evaluation of Analytical Techniques to Predict Viability after Cryopreservation

Verleysen, Hans; Samyn, G.; Bockstaele, Erik Van; Debergh, Pierre

doi:10.1023/b:ticu.0000016483.00158.a9

Cited by 59 publications

(35 citation statements)

References 25 publications

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“…Cell viability tests using 2,3,5-triphenyltetrazolium chloride or fluorescein diacetate frequently produce inaccurate results, due to technical difficulties (Ishikawa et al 1995(Ishikawa et al , 1996Verleysen et al 2004). In this study, the Evans blue exclusion method was effective in determining the viability of BY-2 cells (Figure 2).…”

Section: Discussionmentioning

confidence: 83%

Simple cryopreservation protocol with an encapsulation technique for tobacco BY-2 suspension cell cultures

Kobayashi

Niino

Kobayashi

2005

Plant Biotechnology

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Tobacco BY-2 cells were successfully cryopreserved by a simple slow prefreezing (equilibrium freezing) method using an encapsulation technique. After the cells were immobilized in alginate gel beads, the beads were treated with cryoprotectant solution (2 M glycerol, 0.4 M sucrose) for 45 min. The beads were then transferred to a laboratory freezer at Ϫ30°C, stored for 2 h, and then immersed in liquid nitrogen. To initiate the regrowth of cells, the beads were warmed in a water bath. Following dilution of cryoprotectant solution, the beads were suspended and cultured in normal medium. With this method, suspension cell cultures were regrown within 7 days. There were no differences in the morphology or growth profiles between cryopreserved cell cultures and the original cell cultures.

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Section: Discussionmentioning

confidence: 83%

Simple cryopreservation protocol with an encapsulation technique for tobacco BY-2 suspension cell cultures

Kobayashi

Niino

Kobayashi

2005

Plant Biotechnology

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“…1A and 1D), since the results obtained for viability showed the same trend as those obtained for germination. However, precautions must be taken to interpret the results of this technique since, after recovery from LN, viable plant tissues can still die during regeneration, as well as explants that contain weakened cells, which can be interpreted as non-viable and can regenerate (Verleysen et al, 2004). Due to this fact, the only unquestionable evidence of the success in cryopreservation is germination and normal seedling development after thawing.…”

Section: Resultsmentioning

confidence: 99%

Cryopreservation of Coffea arabica L. Zygotic Embryos by Vitrification

Freitas

Paiva

Sales

et al. 2016

Not Bot Horti Agrobo

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As a consequence of the difficulty in conventional coffee seed storage, biotechnological alternatives such as cryopreservation have been investigated. The objective of this study was to develop a protocol for the cryopreservation of Coffea arabica L. (cv. 'Catuaí Vermelho' -IAC 144) zygotic embryos by vitrification. For the cryopreservation study, the embryos were immersed in Plant Vitrification Solution 2 at different times (0, 10, 25, 50, 100, and 250 min) and two temperatures (0 and 25 °C). Subsequently, the best thawing time was determined in a water bath (1, 3, 5 minutes or directly in Recovery Solution). An anatomical study was conducted on non-stored and stored embryos, with or without the use of Plant Vitrification Solution 2. The immersion in cryoprotectant solution for 100 min at 0 °C allows embryo cryopreservation. Embryos can be directly thawed in Recovery Solution after storage in liquid nitrogen. It was observed that Plant Vitrification Solution 2 reduced internal water content in the cells, allowing subsequent embryo growth resumption.

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“…Seed viability by the TCC test correlated with seed germination test in previous studies, TTC was consecrated as a seed staining solution for general seed cryopreservation research (Verleysen et al, 2004), as well as for vitrified zygotic embryos (Vanda protocorm-like bodies) (Poobathy et al, 2012). It was also useful for orchid seed cryopreservation (Ishikawa et al, 1997;Hirano et al, 2005;Galdiano et al, 2012).…”

Section: Discussionmentioning

confidence: 99%